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Hep-56.1B Cells Europäischer Partner

ArtNr CLS-400202
Hersteller CLS Cell Lines Service
Menge 1 cryovial
Kategorie
Typ Cell line
Zertifikat The certificate of analysis can be requested on the website or via email at info@cytion.com. Please indicate the lot number of your product in the email.
Specific against Mouse (Murine, Mus musculus)
Dry ice Yes
ECLASS 10.1 42040401
ECLASS 11.0 42040401
UNSPSC 41106509
Alias HEP-56.1B, 56.1B, 56.1b
Lieferbar
Manufacturer - Category
Mouse cell lines
Description
The Hep-70.4 hepatoma cell line is derived from a mouse liver tumor, specifically from the C3H/He mouse strain. This cell line is notable for its mutations in the p53 gene, which were identified at different passages during in vitro propagation. At passage number 8, a weak additional signal was detected in the single-strand conformation polymorphism (SSCP) analysis, indicating the presence of a p53 mutation. By passage number 38, two distinct p53 point mutations were identified: a G:C to C:G transversion at codon 135 and a C:G to G:C transversion at codon 138 of exon 5. These mutations led to amino acid changes from alanine to proline and cysteine to tryptophan, respectively.

The Hep-70.4 cell line displays a morphological phenotype that varies significantly during its propagation. Some sublines exhibit an epithelial morphology, while others show a fibroblast-like appearance. This heterogeneity reflects the complex nature of the cell line and its adaptability under different culture conditions. The presence of both normal and mutated p53 alleles in the early passages suggests that the mutations confer a selective growth advantage, leading to the predominance of mutated clones over time.

Intermediate filament protein analysis of the Hep-70.4 cell line revealed the expression of simple keratins K8 and K18, which are typical of normal liver cells, as well as vimentin and keratin K19 to varying degrees. These protein patterns confirm the hepatocytic origin of the cell line and its classification as a hepatoma line. The genomic stability of Hep-70.4 was further assessed through DNA fingerprint analysis, which did not reveal any major structural abnormalities, although changes in the relative intensities of certain bands were observed with increasing passage numbers.
Tissue
Liver
Growth properties
Adherent
Disease
Hepatocellular carcinoma
Age
Adult
Gender
Female
Breed
C57BL/6J
Morphology
Epithelial-like
Biosafety Level
1
Culture Medium
DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 1.5 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a)
Medium Supplements
Supplement the medium with 10% FBS
Subculturing
Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.
Fluid Renewal
Every 3 to 5 days
Freezing Recovery
After thawing, plate the cells at 5 x 10^4 cells/cm^2 and allow the cells to recover from the freezing process and to adhere for at least 24 hours.
Freeze Medium
Handling of Cryopreserved Cultures

Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

Upon receipt, either store the cryovial immediately at temperatures below -150° C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

For immediate culturing, swiftly thaw the vial by immersing it in a 37° C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

Sterility
Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods.
To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.
Safety Precautions
When planning to store a cryovial in liquid nitrogen for future thawing, it is mandatory to adhere to stringent safety measures. Appropriate protective gloves and clothing are essential, and the use of a face mask or safety goggles is required during the transfer of frozen samples to or from the liquid nitrogen tank. This is to mitigate the risk of injury from potential cryovial explosions upon removal, which can result in the projection of sharp fragments.
Disclaimer
Our cells are provided for in vitro laboratory research purposes exclusively and are not intended for clinical or diagnostic use, nor are they to be administered to humans or used for veterinary purposes. Users must adhere to all applicable guidelines and regulations for the handling and use of these cells in a research setting.
Warranty
We stand by the promise of delivering products with high cell viability and robust culture performance. To achieve the best results, please make sure you follow the storage and culture instructions detailed in the product information sheet closely. Your adherence to these guidelines is key to success.
Tumorigenic
Yes, in C57BL/6J mice
Split Ratio
A ratio of 1:4 to 1:8 is recommended
Seeding Density
1 x 10^4 cells/cm^2
Mutational Profile
p53mut (codon 277 in exon 8 =|BiggerAs| Arginin -- Threonin).
Protein Expression
Keratin 8, Keratin 18, Vimentin.
NOTE
Deutsch:
Universitäre Kunden: Für den Erwerb ist ein Material Transfer Agreement oder eine Limited Use Label License auszufüllen.
Kommerzielle Kunden: Für den Erwerb ist ein Material Transfer Agreement oder ein Master Supply Agreement auszufüllen.
Nach eingegangener Bestellung werden ihnen alle relevanten Dokumente zugeschickt.

English:
University Customers: A Material Transfer Agreement or Limited Use Label License must be completed for purchase.
Commercial Customers: A Material Transfer Agreement or Master Supply Agreement must be completed for purchase.
After the order is received, all relevant documents will be sent to you.

Hinweis: Die dargestellten Informationen und Dokumente (Bedienungsanleitung, Produktdatenblatt, Sicherheitsdatenblatt und Analysezertifikat) entsprechen unserem letzten Update und sollten lediglich der Orientierung dienen. Wir übernehmen keine Garantie für die Aktualität. Für spezifische Anforderungen bitten wir Sie, uns eine Anfrage zu stellen.

Alle Produkte sind nur für Forschungszwecke bestimmt. Nicht für den menschlichen, tierärztlichen oder therapeutischen Gebrauch.

Menge: 1 cryovial
Lieferbar: In stock
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